CapZyme-Seq Comprehensively Defines Promoter-Sequence Determinants for RNA 5 ' Capping with NAD(+)

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11320%2F18%3A10389190" target="_blank" >RIV/00216208:11320/18:10389190 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/61388971:_____/18:00490124

Výsledek na webu

<a href="https://doi.org/10.1016/j.molcel.2018.03.014" target="_blank" >https://doi.org/10.1016/j.molcel.2018.03.014</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.molcel.2018.03.014" target="_blank" >10.1016/j.molcel.2018.03.014</a>

Jazyk výsledku

angličtina

Název v původním jazyce

CapZyme-Seq Comprehensively Defines Promoter-Sequence Determinants for RNA 5 ' Capping with NAD(+)

Popis výsledku v původním jazyce

Nucleoside-containing metabolites such asNAD(+) can be incorporated as 50 caps on RNA by serving as non-canonical initiating nucleotides (NCINs) for transcription initiation by RNA polymerase (RNAP). Here, we report CapZyme-seq, a high-throughput-sequencing method that employs NCIN-decapping enzymes NudC and Rai1 to detect and quantify NCIN-capped RNA. By combining CapZyme-seq with multiplexed transcriptomics, we determine efficiencies of NAD(+) capping by Escherichia coli RNAP for similar to 16,000 promoter sequences. The results define preferred transcription start site (TSS) positions for NAD(+) capping and define a consensus promoter sequence for NAD(+) capping: HRRASWW (TSS underlined). By applying CapZyme-seq to E. coli total cellular RNA, we establish that sequence determinants for NCIN capping in vivo match the NAD(+)-capping consensus defined in vitro, and we identify and quantify NCIN-capped small RNAs (sRNAs). Our findings define the promoter-sequence determinants for NCIN capping with NAD(+) and provide a general method for analysis of NCIN capping in vitro and in vivo.

Název v anglickém jazyce

CapZyme-Seq Comprehensively Defines Promoter-Sequence Determinants for RNA 5 ' Capping with NAD(+)

Popis výsledku anglicky

Nucleoside-containing metabolites such asNAD(+) can be incorporated as 50 caps on RNA by serving as non-canonical initiating nucleotides (NCINs) for transcription initiation by RNA polymerase (RNAP). Here, we report CapZyme-seq, a high-throughput-sequencing method that employs NCIN-decapping enzymes NudC and Rai1 to detect and quantify NCIN-capped RNA. By combining CapZyme-seq with multiplexed transcriptomics, we determine efficiencies of NAD(+) capping by Escherichia coli RNAP for similar to 16,000 promoter sequences. The results define preferred transcription start site (TSS) positions for NAD(+) capping and define a consensus promoter sequence for NAD(+) capping: HRRASWW (TSS underlined). By applying CapZyme-seq to E. coli total cellular RNA, we establish that sequence determinants for NCIN capping in vivo match the NAD(+)-capping consensus defined in vitro, and we identify and quantify NCIN-capped small RNAs (sRNAs). Our findings define the promoter-sequence determinants for NCIN capping with NAD(+) and provide a general method for analysis of NCIN capping in vitro and in vivo.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10606 - Microbiology

Projekt

<a href="/cs/project/GA15-05228S" target="_blank" >GA15-05228S: Určení buněčné role HelD, nového vazebného partnera bakteriální RNA polymerázy.</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2018

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Molecular Cell

ISSN

1097-2765

e-ISSN

—

Svazek periodika

70

Číslo periodika v rámci svazku

3

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

12

Strana od-do

553-564

Kód UT WoS článku

000432517300015

EID výsledku v databázi Scopus

2-s2.0-85045536565